Enhanced chemiluminescence ecl western blotting detection reagent

S4 was synthesized as previously described [23 (link)]. 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was purchased from the USB Corporation (Cleveland, OH, USA). L-ascorbic acid, dimethyl sulfoxide, 2′,7′-dichlorofluorescin diacetate (DCFDA), leupeptin, phenylmethylsulfonyl fluoride, and paraformaldehyde were obtained from the Sigma-Aldrich Corporation (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco, Invitrogen (Carlsbad, CA, USA). Enhanced chemiluminescence (ECL)™ Western blotting detection reagent was obtained from Amersham Biosciences (England). ProLong^® Gold antifade reagent with 4′,6-diamidino-2-phenylindole (DAPI) was purchased from Invitrogen (Carlsbad, CA, USA). Mitogen-activated protein (MAP) kinase inhibitors, including c-Jun N-terminal kinases (JNK) inhibitor II, PD98059, and SB203580, were obtained from Calbiochem (Darmstadt, Germany). Tris, sodium dodecyl sulfate (SDS), and Tween 20 were purchased from the USB Corporation (Cleveland, OH, USA). Other reagents and chemicals used in this study were reagent grade.

Type I collagen and phorbol-12, 13-dibutyrate (PDBu) were purchased from Sigma (St Louis, MO). Fura 2-AM was purchased from Molecular Probe (Eugene, OR). The anti-Akt (pan) (40D4) monoclonal antibody (mAb), anti-phospho-Akt (Ser⁴⁷³) polyclonal antibody (pAb), anti-phospho-(Ser) protein kinase C (PKC) substrate pAb, anti-phospho-p38 mitogen-activated protein kinase (MAPK) (Thr¹⁸⁰/Tyr¹⁸²) pAb, anti-p38 MAPK (5F11) mAb, anti-phospho-p44/42 MAPK (ERK1/2) (Thr²⁰²/Tyr²⁰⁴) pAb, anti-p44/42 MAPK (137F5) mAb, anti-phospho-c-Jun N-terminal kinse (JNK) (Thr¹⁸³/Tyr¹⁸⁵) mAb, and anti-JNK pAb were purchased from Cell Signaling (Beverly, MA). The anti-α-tubulin mouse mAb was purchased from Thermo Scientific (Waltham, MA). The Hybond-P polyvinylidene difluoride (PVDF) membrane, an enhanced chemiluminescence (ECL) western blotting detection reagent, a horseradish-peroxidase (HRP)-conjugated donkey anti-rabbit IgG, and a sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, UK).

Anti-HIF-1α and anti-LOX antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-CD11b antibody was purchased from Abcam (Cambridge, UK). Enhanced chemiluminescence (ECL) Western blotting detection reagent was obtained from Amersham (Buckinghamshire, UK). 3,3%-Diaminobenzidine tetrahydrochloride (DAB) immunostaining detection reagent was obtained from Thermo (Runcorn, UK). All other chemicals, including ATP, UTP,γATP, apyrase, and β-aminopropionitrile (βAPN), were purchased from Sigma-Aldrich (St. Louis, MO).